南瓜AFLP反应体系的建立与优化

Establishment and optimization of AFLP reaction system in squash

以印度南瓜矮生等基因系yd为材料,从叶片中提取基因组DNA,对酶切、连接、预扩、选扩反应体系及银染方法进行优化研究,建立高效稳定的南瓜AFLP反应体系,即反应体系为DNA模板量500 ng、37℃酶切5 h、连接3 h、预扩产物稀释50倍及Mg2+1.5 mmoL/L、dNTP 0.19 mmoL/L的效果最好。

An efficient and stable AFLP analysis system was established for squash which was done by selection and optimization in the aspects of DNA extraction, enzyme digestion and ligation, preamplification and selective amplification, silver dying. The results showed that the dosage of template DNA should be 500 ng, the time of digestion should be 5 h at 37℃, the time of ligation should be 3h at 37℃, and the products of preamplification should be diluted to 50 times, the concentration of Mg^2＋ and dNTP should be respectively 1.5 mmoI/L and 0.19 mmoL/L, the result of selective amplification is best.